Expression of the cell adhesion molecules on leukocytes that demarginate during acute maximal exercise

1999 ◽  
Vol 86 (3) ◽  
pp. 970-976 ◽  
Author(s):  
Stephan F. van Eeden ◽  
John Granton ◽  
Jennifer M. Hards ◽  
Barbara Moore ◽  
James C. Hogg
Keyword(s):  
Fluorescence Intensity ◽  
Mean Fluorescence Intensity ◽  
Maximal Exercise ◽  
Flow Cytometric Analysis ◽  
Cycle Ergometer ◽  
Phenotypic Characteristics ◽  
Flow Cytometric ◽  
The Mean ◽  
Low Levels

The pulmonary vascular bed is an important reservoir for the marginated pool of leukocytes that can be mobilized by exercise or catecholamines. This study was designed to determine the phenotypic characteristics of leukocytes that are mobilized into the circulation during exercise. Twenty healthy volunteers performed incremental exercise to exhaustion [maximal O2consumption (V˙o2 max)] on a cycle ergometer. Blood was collected at baseline, at 3-min intervals during exercise, atV˙o2 max, and 30 min after exercise. Total white cell, polymorphonuclear leukocyte (PMN), and lymphocyte counts increased with exercise toV˙o2 max( P < 0.05). Flow cytometric analysis showed that the mean fluorescence intensity of L-selectin on PMN (from 14.9 ± 1 at baseline to 9.5 ± 1.6 atV˙o2 max, P < 0.05) and lymphocytes (from 11.7 ± 1.2 at baseline to 8 ± 0.8 atV˙o2 max, P < 0.05) decreased with exercise. Mean fluorescence intensity of CD11b on PMN increased with exercise (from 10.2 ± 0.6 at baseline to 25 ± 2.5 atV˙o2 max, P < 0.002) but remained unchanged on lymphocytes. Myeloperoxidase levels in PMN did not change with exercise. In vitro studies showed that neither catecholamines nor plasma collected atV˙o2 maxduring exercise changed leukocyte L-selectin or CD11b levels. We conclude that PMN released from the marginated pool during exercise express low levels of L-selectin and high levels of CD11b.

150 α6 Integrin for evaluating sperm quality in bulls with different capacities of invitro embryo production

2020 ◽  
Vol 32 (2) ◽  
pp. 201
Author(s):  
E. G. A. Perez ◽  
D. L. D'Ercole ◽  
M. H. Macedo ◽  
A. A. F. Rodrigues ◽  
R. F. Gonçalves
Keyword(s):  
Bovine Serum Albumin ◽  
Fluorescence Intensity ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Flow Cytometric Analysis ◽  
Fluorescein Isothiocyanate ◽  
Embryo Production ◽  
Logarithmic Amplifier ◽  
Flow Cytometric ◽  
The Mean

The α6 integrin, an adhesion molecule, is expressed on bovine sperm, but major questions about the role of integrins in sperm-oocyte fusion remain unsolved. In this work, we show the results of characterisation of sperm α6 integrin from 4 bulls with different capacities of invitro embryo production using flow cytometric analysis. The bull capacities judged by the rate of blastocyst formation after invitro embryo production with semen from 5 ejaculates per animal were 44.3, 17.1, 13.2, and 15.0% for bulls 1, 2, 3, and 4, respectively (P&lt;0.05). For flow cytometric analysis, surface expression of α6 integrin was evaluated using a fluorescence-activated cell sorter (FACScan, Coulter Electronics) using a 520-nm excitation from an argon laser at 150 mW for excitation. Frozen-thawed sperm were centrifuged at 700×g for 10min and washed once in warm phosphate buffered saline (PBS). Briefly, the spermatozoa (5×105 cells per sample) were resuspended in 100μL of PBS containing 1% bovine serum albumin. After washing 3 times, the live cells were incubated at room temperature for 1h with 100μL (1:500) of α6 monoclonal antibody (Chemicon) in PBS (0.1M, pH 7.4) with 1% bovine serum albumin. After washing three times, the cells were incubated at 4°C for 1h in the dark with the fluorescein isothiocyanate-conjugated F(ab)2 fragment of affinity isolated goat anti-mouse antibody (Invitrogen). The cells were washed three times, resuspended in 100μL of PBS, and analysed. For each sample, 10 000 cells were recorded at a flow rate of 200-300 cells s−1 using forward scatter (cell size) and side angle of light scatter (cell density), the first using a logarithmic amplifier and the second using a linear amplifier. The fluorescence data were collected using the logarithmic amplifier. The percentage of positive cells and the mean fluorescence channel on a 1023-channel scale were calculated using Epics Profile II software (Epics II Software). To define the forward and side-scatter regions corresponding to sperm, binding of fluorescein isothiocyanate-conjugated Pisum sativum agglutinin to the acrosome was used for setting the bitmap on the dot plot. Initially, the percentage of spermatozoa stained with α6 antibody was calculated on a per-individual basis. Subsequently, the mean±standard deviation was calculated for each group. The Mann-Whitney U test was used to compare the differences in the expression of α6 between groups. Expression of α6 integrin was higher in bull 1 (control) than in bulls with low capacities of invitro embryo production. The spermatozoa from bull 1 was distributed in a single broad peak with a mean fluorescence intensity of 36.16±4.17%. The increased distance of the fluorescence intensity in bull 1 compared with the weak fluorescence peaks in the others bulls reflects the increased width and distance of the population distribution. In conclusion, α6 integrin may be used as a biomarker to evaluate sperm quality. This study was supported by FAPESP grants 2010/01077-9, 2011/18085-7, and 2016/00976-6.

scholarly journals Flow Cytometric Analysis of T, B, and NK Cells Antigens in Patients with Mycosis Fungoides

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Serkan Yazıcı ◽  
Emel Bülbül Başkan ◽  
Ferah Budak ◽  
Barbaros Oral ◽  
Şaduman Balaban Adim ◽  
...  
Keyword(s):  
Cell Surface ◽  
Mycosis Fungoides ◽  
Peripheral Blood ◽  
Prognostic Value ◽  
Flow Cytometric Analysis ◽  
Surface Antigens ◽  
Cell Surface Antigens ◽  
Flow Cytometric ◽  
Hla Dr ◽  
The Mean

We retrospectively analyzed the clinicopathological correlation and prognostic value of cell surface antigens expressed by peripheral blood mononuclear cells in patients with mycosis fungoides (MF). 121 consecutive MF patients were included in this study. All patients had peripheral blood flow cytometry as part of their first visit. TNMB and histopathological staging of the cases were retrospectively performed in accordance with International Society for Cutaneous Lymphomas/European Organization of Research and Treatment of Cancer (ISCL/EORTC) criteria at the time of flow cytometry sampling. To determine prognostic value of cell surface antigens, cases were divided into two groups as stable and progressive disease. 17 flow cytometric analyses of 17 parapsoriasis (PP) and 11 analyses of 11 benign erythrodermic patients were included as control groups. Fluorescent labeled monoclonal antibodies were used to detect cell surface antigens: T cells (CD3+, CD4+, CD8+, TCRαβ+, TCRγδ+, CD7+, CD4+CD7+, CD4+CD7−, and CD71+), B cells (HLA-DR+, CD19+, and HLA-DR+CD19+), NKT cells (CD3+CD16+CD56+), and NK cells (CD3−CD16+CD56+). The mean value of all cell surface antigens was not statistically significant between parapsoriasis and MF groups. Along with an increase in cases of MF stage statistically significant difference was found between the mean values of cell surface antigens. Flow cytometric analysis of peripheral blood cell surface antigens in patients with mycosis fungoides may contribute to predicting disease stage and progression.

New thiazacridine agents: Synthesis, physical and chemical characterization, and in vitro anticancer evaluation

2016 ◽  
Vol 36 (10) ◽  
pp. 1059-1070 ◽  
Author(s):  
MBO Chagas ◽  
NCC Cordeiro ◽  
KMR Marques ◽  
MG Rocha Pitta ◽  
MJBM Rêgo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cytotoxic Activity ◽  
Cell Cycle Progression ◽  
Antitumor Agents ◽  
Flow Cytometric Analysis ◽  
Cell Lineage ◽  
Chemical Characterization ◽  
Flow Cytometric ◽  
Physical And Chemical

A series of new thiazacridine agents were synthesized and evaluated as antitumor agents, in terms of not only their cytotoxicity but also their selectivity. The cytotoxicity assay confirmed that all compounds showed cytotoxic activity and selectivity. The new compound, 3-acridin-9-ylmethyl-5-(5-bromo-1 H-indol-3-ylmethylene)-thiazolidine-2,4-dione (LPSF/AA29 – 7a), proved to be the most promising compound as it presents lower half-maximal inhibitory concentration (IC50) values (ranging from 0.25 to 68.03 µM) depending on cell lineage. In HepG2 cells, the lowest IC50 value was exhibited by 3-acridin-9-ylmethyl-5-(4-piperidin-1-yl-benzylidene)-thiazolidine-2,4-dione (LPSF/AA36 – 7b; 46.95 µM). None of the synthesized compounds showed cytotoxic activity against normal cells (IC50 > 100 µM). The mechanism of death induction and cell cycle effects was also evaluated. Flow cytometric analysis revealed that the compounds LPSF/AA29 – 7a and LPSF/AA36 – 7b significantly increased the percentage of apoptotic cells and induced G2/M arrest in the cell cycle progression. Therefore, these new thiazacridine derivatives constitute promising antitumor agents whose cytotoxicity and selectivity properties indicate they have potential to contribute to or serve as a basis for the development of new cancer drugs in the future.

Effects of high-intensity cycle exercise on sympathoadrenal-medullary response patterns

1991 ◽  
Vol 70 (1) ◽  
pp. 8-14 ◽  
Author(s):  
W. J. Kraemer ◽  
J. F. Patton ◽  
H. G. Knuttgen ◽  
C. J. Hannan ◽  
T. Kettler ◽  
...  
Keyword(s):  
High Intensity ◽  
Maximal Exercise ◽  
Cycle Ergometer ◽  
Plasma Norepinephrine ◽  
Response Patterns ◽  
O2 Uptake ◽  
Cycle Exercise ◽  
Before And After ◽  
The Mean ◽  
The Individual

Plasma proenkephalin peptide F immunoreactivity and catecholamines were examined on separate days in nine healthy males before and after maximal exercise to exhaustion at four intensities [36, 55, 73, and 100% of maximal leg power (MLP)] by use of a computerized cycle ergometer. The mean duration of 36, 55, 73, and 100% MLP was 3.31, 0.781, 0.270, and 0.1 min, respectively. All intensities were greater than those eliciting peak O2 uptake for the individual subjects. Blood samples were obtained before, immediately after exercise, and 5 and 15 min after exercise. Significant (P less than 0.05) increases in plasma peptide F immunoreactivity (i.e., from mean resting value of 0.18 to 0.43 pmol/ml) were observed immediately after exercise at 36% MLP. Significant increases in plasma epinephrine were observed immediately after exercise at 36% MLP (i.e., from mean resting value of 2.22 to 3.11 pmol/ml) and 55% MLP (i.e., from mean resting value of 1.67 to 2.98 pmol/ml) and 15 min after exercise at 100% MLP (i.e., from mean resting value of 1.92 to 3.88 pmol/ml). Significant increases for plasma norepinephrine were observed immediately after exercise (36, 55, 73, and 100% MLP), 5 min after exercise (36, 55, and 73% MLP), and 15 min after exercise (36% MLP). Increases in whole blood lactate were observed at all points after exercise for 36, 55, and 73% MLP and 5 min after exercise for 100% MLP. These data show that brief high-intensity exercise results in differential response patterns of catecholamines and proenkephalin peptide F immunoreactivity.

Exploration of cytotoxic and genotoxic endpoints following sub-chronic oral exposure to titanium dioxide nanoparticles

2019 ◽  
Vol 35 (9) ◽  
pp. 577-592 ◽  
Author(s):  
Srijita Chakrabarti ◽  
Danswrang Goyary ◽  
Sanjeev Karmakar ◽  
Pronobesh Chattopadhyay
Keyword(s):  
Titanium Dioxide ◽  
Titanium Dioxide Nanoparticles ◽  
Flow Cytometric Analysis ◽  
Raw 264.7 Cells ◽  
Oral Exposure ◽  
Chromosomal Breakage ◽  
Flow Cytometric ◽  
Higher Doses

Health hazards of titanium dioxide nanoparticles (TiO2-NPs) have raised severe concerns because of the paucity of information regarding the toxic effects among the population. In the present research, the in vitro and in vivo cytotoxic potential of TiO2-NPs were evaluated using flow cytometric techniques. Further, in vitro and in vivo genotoxic endpoints were estimated by means of comet, micronucleus (MN), and chromosomal aberration (CA) assays. In vitro analysis was performed at the concentration range of 10–100 µg/mL using murine RAW 264.7 cells. In vivo experiments were conducted on Albino mice (M/F) by exposing them to 200 and 500 mg/kg TiO2-NPs for 90 days. Decreased percentage of cell viability with higher doses of TiO2-NPs was evident in both in vitro and in vivo flow cytometric analysis. Further, an impaired cell cycle (G0/G1, S, and G2/M) was reflected in the present investigation following the exposure to TiO2-NPs. Increased comet scores such as tail length, % DNA in tail, tail moment, and olive moment were also observed with the higher doses of TiO2-NPs in vitro and in vivo comet assays. Finally, the in vivo MN and CA assays revealed the formation of MN and chromosomal breakage following the exposure to TiO2-NPs.

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